Title of Practical: Identification of Biological Chemicals

Aim: To identify the various biological chemicals present in five unknown substances.
Apparatus: Test tubes, test tube rack, unknown solutions 1-5,
Procedure:
The results were reported in table format

The test for starch (Iodine Test)

1. Five test-tubes were labelled
appropriately, one for each of the five The test for non-reducing sugar (Benedict's
unknown solutions Test)
2. 2 cm3 of each solution was poured into 1. A beaker was half filled with tap water
the appropriately labelled test-tube. and was placed on a tripod and gauze.
3. To each test-tube, using a dropper, The water was heated with a Bunsen
three drops of iodine solution were burner.
added, 2. The five test-tubes were labelled
4. The test-tubes were shaken to mix the appropriately, one for each of the five
contents. unknown solutions.
5. Any colour changes apart from the 3. 2 cm3 of each solution was poured into
yellow colour of iodine itself were the appropriately labelled test-tube.
observed. 4. Three drops of dilute hydrochloric acid
The results were recorded in an were added.
appropriate table. 5. The test-tubes were placed in the
beaker of hot water and the mixture
was boiled for 2 minutes.
The test for reducing sugars (Benedict's Test) 6. The test tube was removed and was
1. A beaker was half filled with tap water allowed to cool to room temperature.
and placed on a tripod and gauze. The 7. Three drops of dilute sodium hydrogen
water was heated with a Bunsen carbonate were added, to neutralize
burner. the acid (the mixture is neutral when
2. Five test-tubes were labelled fizzing stops).
appropriately, one for each of the five 8. 1cm3 of Benedict’s solution was added
unknown solutions to each test tube.
3. 2 cm3 of each solution was poured into 9. The test-tubes were placed in the
the appropriately labelled test-tube. beaker of hot water, and the flame was
4. 1cm3 of Benedict’s solution was added adjusted to keep the water just boiling.
to each test tube. 10. After about 5 minutes, the flame was
5. The test-tubes were placed in a beaker extinguished.
of hot water, and the flame was 11. The five test tubes were placed in a
adjusted to keep the water just boiling. test-tube rack and compare the colours.
6. After about 5 minutes, the flame was The resulting colours were recorded
extinguished.
The five test tubes were placed in a test-
tube rack and the colours were compared.
The resulting colours were recorded

Lab. #: 1
Skill: Observation, Recording, Reporting (ORR) Manipulation and Measurement (MM)
Syllabus Topic: Aspects of Biochemistry
The test for Protein (Biuret Test) Test for Lipids (Emulsion Test)

1. Five test-tubes were labelled 1. Five test-tubes appropriately, one for

appropriately, one for each of the five each of the five unknown solutions
unknown solutions 2. Pour 2 cm3 of each solution into the
2. 2 cm3 of each solution was poured into appropriately labelled test-tube.
the appropriately labelled test-tube. 3. About 2 cm3 of ethanol was added to
3. An equal volume of dilute (5%) each test tube.
potassium hydroxide solution was 4. The contents of the test-tubes were
added to each test-tube shaken to dissolve the lipid in the
4. The test-tubes were shaken to mix the ethanol.
contents 5. About 2 cm3 of water was added to
5. Three drops of 1% copper sulphate each test tube.
solution were added using a dropper. 6. The resulting colours were recorded.
6. The test-tubes were shaken to mix the
contents.
7. The test tubes were returned to the
rack, and were left for ten seconds
before the resulting colours were
recorded.

Lab. #: 1
Skill: Observation, Recording, Reporting (ORR) Manipulation and Measurement (MM)
Syllabus Topic: Aspects of Biochemistry
Observations: Test Tube A

Table showing the type of test, aim, method observation and inferences for test tube A

Observation
Test Aim Method Inferences
Initial Final
Starch To determine the 1. Three drops of iodine The colour The colour Starch is present
presence of starch solution were added, using of the of the
using Iodine a dropper solution is solution
Solution 2. The test-tube was then colourless. becomes
shaken to mix the Blue-
contents. Black.
3. Any colour changes apart
from the yellow colour of
iodine itself were
observed.
4. The results were recorded
in this table.
Reducing Sugar To determine the 1. 1 cm3 of Benedict’s solution The colour The colour Reducing Sugars
presence of was added to test tube. of the of the are not present
reducing sugars 2. The test-tube was placed in solution is solution
using Benedict’s a beaker of hot water, and colourless. becomes
Solution the flame was adjusted to light blue.
keep the water just boiling.
3. After about 5 minutes, the
test tube was removed
from the hot water bath
4. The test tube was placed in
a rack and the colours were
observed and recorded.

1
Test Tube A
Observations: Test Tube A

Non-reducing Sugar To determine the 1. Three drops of dilute The colour The colour Non-reducing
presence of non- hydrochloric acid were of the of the sugar is not
reducing sugars added to the test tube. solution is solution present
using Dilute 2. The test-tube was placed in colourless. becomes
Hydrochloric Acid a beaker of hot water and light blue
and Benedict’s the mixture was boiled for then
Solution becomes
2 minutes.
green.
3. The test tube was removed
and allowed to cool to
room temperature.
4. Three drops of dilute
sodium hydrogen
carbonate were added to
neutralize the acid.
5. 1 cm3 of Benedict’s solution
was added to the test tube.
6. The test-tube was placed in
a beaker of hot water, and
the flame was adjusted to
keep the water just boiling.
7. After about 5 minutes, the
test tube was removed
from the hot water bath.
8. The test tube was then
placed in a rack and the
colours were observed and
recorded.

2
Test Tube A
Observations: Test Tube A

Protein To determine the 1. An equal volume of dilute The colour The colour Protein is not
presence of protein (5%) potassium hydroxide of the of the present
using Biuret Solution solution was added to the solution is solution
test-tube colourless. became
2. The test-tube was shaken Blue
to mix the contents
3. Three drops of 1% copper
sulphate solution was
added using a dropper.
4. The test-tube was shaken
to mix the contents.
5. The test tube was then
placed in a rack, and the
colours were observed and
recorded after three
seconds had past.

Lipids To determine the 1. About 2 cm3 of ethanol was The colour The colour Lipids are not
presence of lipids added to the test tube. of the of the present
using ethanol 2. The contents of the test- solution is solution is
tube was shaken to dissolve colourless. colourless.
the lipid in the ethanol.
3. about 2 cm3 of water was
added to the test tube.
4. The test tube was then
placed in a rack, and the
colours were observed and
recorded

3
Test Tube A
Observations: Test Tube B

Table showing the type of test, aim, method observation and inferences for test tube B

Observation
Test Aim Method Inferences
Initial Final
Starch To determine 1. Three drops of iodine solution The colour The colour Starch is not
the presence were added, using a dropper of the of the present
of starch using 2. The test-tube was then shaken to solution is solution
Iodine mix the contents. colourless. becomes
Solution 3. Any colour changes apart from yellow.
the yellow colour of iodine itself
were observed.
4. The results were recorded in this
table.
Reducing Sugar To determine 1. 1 cm3 of Benedict’s solution was The colour The colour Reducing sugars
the presence added to test tube. of the of the are not present
of reducing 2. The test-tube was placed in a solution is solution
sugars using beaker of hot water, and the colourless. becomes
Benedict’s flame was adjusted to keep the light blue.
Solution water just boiling.
3. After about 5 minutes, the test
tube was removed from the hot
water bath
4. The test tube was placed in a
rack and the colours were
observed and recorded.

1
Test Tube B
Observations: Test Tube B

Non-reducing Sugar To determine 1. Three drops of dilute The colour The colour A high
the presence hydrochloric acid were added to of the of the concentration of
of non- the test tube. solution is solution non-reducing
reducing 2. The test-tube was placed in a colourless. becomes sugars is
sugars using beaker of hot water and the light blue, present.
Dilute mixture was boiled for 2 minutes. then
Hydrochloric becomes
3. The test tube was removed and
Acid and Cloudy
allowed to cool to room
Benedict’s Orange.
Solution temperature.
4. Three drops of dilute sodium
hydrogen carbonate were added
to neutralize the acid.
5. 1 cm3 of Benedict’s solution was
added to the test tube.
6. The test-tube was placed in a
beaker of hot water, and the
flame was adjusted to keep the
water just boiling.
7. After about 5 minutes, the test
tube was removed from the hot
water bath.
8. The test tube was then placed in
a rack and the colours were
observed and recorded.

2
Test Tube B
Observations: Test Tube B

Protein To determine 1. An equal volume of dilute The colour The colour Protein is not
the presence (5%) potassium hydroxide of the of the present
of protein solution was added to the solution is solution
using Biuret test-tube colourless. becomes
Solution 2. The test-tube was shaken to blue.
mix the contents
3. Three drops of 1% copper
sulphate solution was added
using a dropper.
4. The test-tube was shaken to
mix the contents.
5. The test tube was then
placed in a rack, and the
colours were observed and
recorded after three seconds
had past.
Lipids To determine 1. About 2 cm3 of ethanol was The colour The colour Lipids are not
the presence added to the test tube. of the of the present
of lipids using 2. The contents of the test-tube was solution is solution is
ethanol shaken to dissolve the lipid in the colourless. colourless.
ethanol.
3. about 2 cm3 of water was added
to the test tube.
4. The test tube was then placed in a
rack, and the colours were
observed and recorded

3
Test Tube B
Observations: Test Tube C

Table showing the type of test, aim, method observation and inferences for test tube C

Observation
Test Aim Method Inferences
Initial Final
Starch To determine 1. Three drops of iodine solution Colourless Yellow Starch is not
the presence were added, using a dropper present
of starch 2. The test-tube was then shaken
using Iodine to mix the contents.
Solution 3. Any colour changes apart from
the yellow colour of iodine
itself were observed.
4. The results were recorded in
this table.

Reducing Sugar To determine 1. 1 cm3 of Benedict’s solution Colourless Brick Red A very high
the presence was added to test tube. concentration
of reducing 2. The test-tube was placed in a of reducing
sugars using beaker of hot water, and the sugars are
Benedict’s flame was adjusted to keep the present
Solution water just boiling.
3. After about 5 minutes, the test
tube was removed from the
hot water bath
4. The test tube was placed in a
rack and the colours were
observed and recorded.

1
Test Tube C
Observations: Test Tube C

Non-reducing To determine 1. Three drops of dilute Colourless Brick Red The sugars
Sugar the presence hydrochloric acid were added present are
of non- to the test tube. reducing, as
reducing 2. The test-tube was placed in a the test prior to
sugars using beaker of hot water and the this responded
Dilute mixture was boiled for 2 positive as well.
Hydrochloric
minutes.
Acid and
3. The test tube was removed and
Benedict’s
Solution allowed to cool to room
temperature.
4. Three drops of dilute sodium
hydrogen carbonate were
added to neutralize the acid.
5. 1 cm3 of Benedict’s solution
was added to the test tube.
6. The test-tube was placed in a
beaker of hot water, and the
flame was adjusted to keep the
water just boiling.
7. After about 5 minutes, the test
tube was removed from the
hot water bath.
8. The test tube was then placed
in a rack and the colours were
observed and recorded.

Protein To determine 1. An equal volume of dilute (5%) The colour of The colour of Protein is not

2
Test Tube C
Observations: Test Tube C

the presence potassium hydroxide solution the solution is the solution present
of protein was added to the test-tube colourless. becomes
using Biuret 2. The test-tube was shaken to blue.
Solution mix the contents
3. Three drops of 1% copper
sulphate solution was added
using a dropper.
4. The test-tube was shaken to
mix the contents.
5. The test tube was then placed
in a rack, and the colours were
observed and recorded after
three seconds had past.
Lipids To determine 5. About 2 cm3 of ethanol was The colour of The colour of Lipids is not
the presence added to the test tube. the solution is the solution present
of lipids using 6. The contents of the test-tube colourless. is colourless.
ethanol were shaken to dissolve the lipid
in the ethanol.
7. About 2 cm3 of water was
added to the test tube.
8. The test tube was then placed in
a rack, and the colours were
observed and recorded

3
Test Tube C
Observations: Test Tube D

Table showing the type of test, aim, method observation and inferences for test tube D

Observation
Test Aim Method Inferences
Initial Final
Starch To determine 1. Three drops of iodine solution The colour The colour Starch is not
the presence were added, using a dropper of the of the present
of starch 2. The test-tube was then shaken to solution is solution
using Iodine mix the contents. golden remains
Solution 3. Any colour changes apart from yellow golden
the yellow colour of iodine itself yellow
with a
were observed.
yellow
4. The results were recorded in this
precipitat
table. e floating
at the top
Reducing Sugar To determine 1. 1 cm3 of Benedict’s solution was The colour The colour Reducing Sugars
the presence added to test tube. of the of the are not present
of reducing 2. The test-tube was placed in a solution is solution
sugars using beaker of hot water, and the golden remains
Benedict’s flame was adjusted to keep the yellow golden
Solution water just boiling. yellow
with a
3. After about 5 minutes, the test
light blue
tube was removed from the hot
precipitat
water bath e floating
4. The test tube was placed in a rack at the top
and the colours were observed
and recorded.

1
Test Tube D
Observations: Test Tube D

Non-reducing Sugar To determine 1. Three drops of dilute hydrochloric The colour The colour Non-reducing
the presence acid were added to the test tube. of the of the sugars are not
of non- 2. The test-tube was placed in a solution is solution present
reducing beaker of hot water and the golden remains
sugars using mixture was boiled for 2 minutes. yellow golden
Dilute 3. The test tube was removed and yellow
Hydrochloric with a
allowed to cool to room
Acid and light blue
temperature.
Benedict’s precipitat
Solution 4. Three drops of dilute sodium e floating
hydrogen carbonate were added at the top
to neutralize the acid.
5. 1 cm3 of Benedict’s solution was
added to the test tube.
6. The test-tube was placed in a
beaker of hot water, and the
flame was adjusted to keep the
water just boiling.
7. After about 5 minutes, the test
tube was removed from the hot
water bath.
8. The test tube was then placed in a
rack and the colours were
observed and recorded.

2
Test Tube D
Observations: Test Tube D

Protein To determine 1. An equal volume of dilute (5%) The colour The colour Protein is not
the presence potassium hydroxide solution was of the of the present
of protein added to the test-tube solution is solution
using Biuret 2. The test-tube was shaken to mix golden remains
Solution the contents yellow golden
3. Three drops of 1% copper yellow
with a
sulphate solution was added
blue
using a dropper.
precipitat
4. The test-tube was shaken to mix e floating
the contents. at the top
5. The test tube was then placed in a
rack, and the colours were
observed and recorded after
three seconds had past.
Lipids To determine 1. About 2 cm3 of ethanol was added The colour The colour Lipids are
the presence to the test tube. of the of the present
of lipids using 2. The contents of the test-tube was solution is solution
ethanol shaken to dissolve the lipid in the golden becomes
ethanol. yellow milky
3. About 2 cm3 of water was added white
to the test tube.
4. The test tube was then placed in a
rack, and the colours were
observed and recorded

3
Test Tube D
Observations: Test Tube E

Table showing the type of test, aim, method observation and inferences for test tube E

Observation
Test Aim Method Inferences
Initial Final
Starch To determine 1. Three drops of iodine solution Cloudy Yellow Starch is not
the presence of were added, using a dropper White present
starch using 2. The test-tube was then shaken
Iodine Solution to mix the contents.
3. Any colour changes apart from
the yellow colour of iodine
itself were observed.
4. The results were recorded in
this table.
Reducing Sugar To determine 5. 1 cm3 of Benedict’s solution Cloudy Light Blue Reducing sugars
the presence of was added to test tube. White are not present
reducing sugars 6. The test-tube was placed in a
using Benedict’s beaker of hot water, and the
Solution flame was adjusted to keep
the water just boiling.
7. After about 5 minutes, the test
tube was removed from the
hot water bath
8. The test tube was placed in a
rack and the colours were
observed and recorded.

1
Test Tube E
Observations: Test Tube E

Non-reducing Sugar To determine 1. Three drops of dilute The colour The colour Non-reducing
the presence of hydrochloric acid were added of the of the sugars are not
non-reducing to the test tube. solution is solution present
sugars using 2. The test-tube was placed in a cloudy remains
Dilute beaker of hot water and the white cloudy
Hydrochloric mixture was boiled for 2 white with
Acid and a light blue
minutes.
Benedict’s precipitate
3. The test tube was removed
Solution floating at
and allowed to cool to room the top
temperature.
4. Three drops of dilute sodium
hydrogen carbonate were
added to neutralize the acid.
5. 1 cm3 of Benedict’s solution
was added to the test tube.
6. The test-tube was placed in a
beaker of hot water, and the
flame was adjusted to keep
the water just boiling.
7. After about 5 minutes, the test
tube was removed from the
hot water bath.
8. The test tube was then placed
in a rack and the colours were
observed and recorded.

2
Test Tube E
Observations: Test Tube E

Protein To determine 1. An equal volume of dilute (5%) The colour The colour Protein is
the presence of potassium hydroxide solution of the of the present
protein using was added to the test-tube solution is solution
Biuret Solution 2. The test-tube was shaken to cloudy becomes
mix the contents white purple
3. Three drops of 1% copper
sulphate solution was added
using a dropper.
4. The test-tube was shaken to
mix the contents.
5. The test tube was then placed
in a rack, and the colours were
observed and recorded after
three seconds had past.
Lipids To determine 1. About 2 cm3 of ethanol was The colour The colour Lipids are not
the presence of added to the test tube. of the of the present
lipids using 2. The contents of the test-tube solution is solution
ethanol was shaken to dissolve the cloudy remains
lipid in the ethanol. white cloudy
3. About 2 cm3 of water was white
added to the test tube.
4. The test tube was then placed
in a rack, and the colours were
observed and recorded

3
Test Tube E
Lab #1

Discussion:

Conclusion: Based on the results, it is apparent that only the contents of test tube A contained starch.
Both, test tube ‘A’ and ‘B’ contained non-reducing sugars. Only test tube ‘C’ contained reducing sugars.
Only test tube ‘D’ contained lipids, and test tube ‘E’ contained protein.
Conduct a quantitative study to determine the protein content of different milk powder brands.

Experiment: 2
Skill: Observation, Recording, Reporting (ORR)
Date:
Syllabus Topic: Aspects of Biochemistry
Title of Practical: Quantitative Analysis of Starch
Aim: To investigate and compare quantitatively starch
Apparatus: Microscope, scalpel, onion bulb, forceps, 0.05M Iodine, microscope slide, cover slip,
mounted needle, pencil, paper, rule and scale slide, 1% starch solution, 5 test tubes, test tube rack, 10
cm3 pipette or syringe, distilled water
Procedure
a serial dilution of starch solution was made as follows:
1. five clean test-tubes were placed in a rack and were labelled 1-5.
2. 10 cm3 of water was measured using a syringe and was placed in the 1% starch solution in test
tube 1.
3. 1 cm3 of the solution from test tube 1 was taken out with the syringe and was placed in test
tube 2.
4. 9 cm3 of water was added with the syringe to test tube 2 and was shaken sideways to mix the
contents.
5. The 1 cm3 of 1% starch solution was now diluted ten times to make a 0.1 % solution.
6. 1 cm3 of this 0.1% starch solution was transferred from test tube 2 into test tube 3.
7. 9 cm3 of water was added to test tube 3 and shaken. This would give a 0.01 % solution.
8. 1 cm3 of the solution was transferred from tube 3 to tube 4 and was diluted with 9 cm 3 of water
making a .001% solution.
9. the operation for test tube 5.was repeated
10. Test tubes 1-4 would now have had 9 cm3 of solution in them but test tube 5 would have 10
cm3 so 1 cm3 of this solution from tube 5 so that the subsequent test is a fair one.
11. Four drops of iodine solution were added to each tube and was shaken to mix.
12. piece of onion tissue was placed in each test tube and was observed under microscope.
13. the presence of starch in onion tissue was noted
14. a white paper was placed behind the test tube and record colour development in a table
Brown (-) Blue-Black (++++)

1
Conduct a quantitative study to determine the protein content of different milk powder brands.

Observation
Table showing…
Concentration of Volume of Solution Observation Colour Intensity
Solution

1
Conduct a quantitative study to determine the protein content of different milk powder brands.

Discussion
1. What is the least concentration of starch that iodine solution will detect? Qualify your answer.
2. Do you think a .0001 % solution contains any starch at all?
Conclusion